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Phosphorylation State of Tyrosine Hydroxylase in the Stalk-Median Eminence Is Decreased by Progesterone in Cycling Female Rats

Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois 62901

Address all correspondence and requests for reprints to: Dr. Lydia A. Arbogast, Department of Physiology, Southern Illinois University School of Medicine, Carbondale, Illinois 62901-6523. E-mail: larbogast{at}siumed.edu.

Progesterone has the capacity to suppress hypothalamic dopaminergic neuronal activity on proestrous afternoon and prolong or amplify the preovulatory prolactin surge in rats. In the present study, we examined enzyme activity and phosphorylation state of tyrosine hydroxylase (TH) in the stalk-median eminence of cycling female rats on proestrus and estrus and related these to circulating progesterone levels. Phospho-TH levels were evaluated by Western blot analysis. TH activity was determined from the rate of 3,4-dihydroxyphenylalanine (DOPA) accumulation. Phospho-TH levels at Ser-19, Ser-31, and Ser-40 were similar at 1100, 1300, and 1500 h on proestrus but declined at 1700, 1900, and 2200 h, coincident with rising serum progesterone levels. Similarly, DOPA accumulation was 30–50% lower at 1700, 1900, and 2200 h as compared with 1100–1500 h on proestrus. Ser-31 and Ser-40 phosphorylation states were increased by 1100 h on estrus to a level similar to 1100 h on proestrus, whereas DOPA accumulation was 30% greater on estrous as compared with proestrous morning. There were no significant differences among the several time points on proestrus and estrus with regard to TH protein or β-tubulin levels. Exogenous progesterone administration (7.5 mg/kg, sc) before the preovulatory progesterone surge decreased TH activity and phospho-TH at Ser-19, Ser-31, and Ser-40, accompanied by premature increased serum prolactin. Our study suggests that decreased TH phosphorylation at Ser-19, Ser-31, and Ser-40 contributes to the decline in TH activity in the stalk-median eminence on proestrous afternoon and that progesterone may cause this initial cytoplasmic response of TH dephosphorylation.